PROJECT SUMMARY/ABSTRACT This proposal requests funds to acquire a BIOTEK Cytation 5 multimode (imaging) reader with BioSpa automated incubator and MultiFlo liquid handler for the Medical University of South Carolina to support the drug discovery effort of NIH-funded investigators. The equipment will be housed in the Department of Drug Discovery & Biomedical Sciences (DDBS) and will be utilized by the Cell & Molecular Imaging Core (CMI) and the Drug Discovery Center (DDC) to promote scientific discovery and speed translation of basic scientific findings into preclinical and clinical studies through drug discovery. The DDC also facilitates the discovery of new chemical probes and provides students the opportunity to learn innovative techniques while doing real world experimentation in drug discovery. These educational opportunities are critical to the mission of MUSC and the DDBS. CMI Investigators currently utilize available confocal microscopes (Zeiss LSM 880 with Airyscan, Zeiss LSM 510 and Olympus Fv10i) to visualize changes in cellular morphology, bioenergetics, proliferation and protein trafficking in 8-12-well chamber slides. While the Zeiss LSM 880 system is equipped with an incubation chamber for 96-well plates, this chamber can only be used with objectives 10x or lower, and neither the speed of imaging nor the software are appropriate for high content analysis. Further, the Zeiss LSM 880 is utilized almost 24/7 for various other projects. The Olympus Fv10i confocal microscope also has automated imaging capabilities. However, the Olympus 10i can only handle either 3x 35 mm dishes or one chamber slide at a time. The DDC previously utilized an InCell 2000 high-content analyzer (GE HealthCare) for drug discovery projects originally purchased in 2008 but currently is using a Hermes imaging system from IDA Biomedical acquired in 2011. Unfortunately, both systems are obsolete. The InCell2000 analyzer is no longer operable, and per the manufacturer it would not be cost effective to repair it. This older model Hermes system can image only 3 different fluorescence channels, which limits the users? capability of multiplexing. Further, the environmental chamber does not accommodate all standard plates for long-term live cell analysis and has a limited range of temperature and CO2 control and no O2 control. Moreover, the Hermes cannot be upgraded with robotics. In summary, currently available systems at the CMI are not suitable for higher throughput imaging of signaling pathways, cellular interactions and metabolic changes. Further, the high-content analyzers in the Drug Discovery Center (DDC) are obsolete, have limited detection capabilities and cannot conduct high-throughput screens using phenotypic assays that require the simultaneous measurement of multiple readout parameters. The acquisition of a BIOTEK Cytation 5 multimode (imaging) reader with BioSpa automated incubator and MultiFlo liquid handler would greatly expand the screening capabilities of both the CMI and DDC and rapidly advance drug discovery projects being conducted by MUSC faculty.